LOCUS       sequence-176573-        5666 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express vaccinia virus D12L in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5666)
  AUTHORS   Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N, Onishi M, 
            Matsuura Y, Taniguchi K, Kobayashi T
  TITLE     Entirely plasmid-based reverse genetics system for rotaviruses.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 
            10.1073/pnas.1618424114.
  PUBMED    28137864
REFERENCE   2  (bases 1 to 5666)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 
            10.1073/pnas.1618424114."
COMMENT     Sequence Label: pCAG-D12L
FEATURES             Location/Qualifiers
     source          1..5666
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     215..237
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     299..318
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1341..1360)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1406..1461
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1460..1479)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1813..1833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1818..2147
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1998..2133
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2240..2839
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2240..2259)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2736..2756
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3069..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3106..3125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3160..3179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3271..3288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3442..4030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3522..3541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4201..4992,4993..5061))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4824..4843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5062..5166)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5197..5576
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
       61 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
      121 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
      181 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
      241 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
      301 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattaattaa gtttaaacgg
      361 cgcgccgata tcttaagatc tatggatgaa attgtaaaaa atatccggga gggaacgcat
      421 gtccttcttc cattttatga aacattgcca gaacttaatc tgtctctagg taaaagccca
      481 ttacctagtc tggaatacgg agctaattac tttcttcaga tttctagagt taatgatcta
      541 aatagaatgc cgaccgacat gttaaaactt tttacacatg atatcatgtt accagaaagc
      601 gatctagata aagtctatga aattttaaag attaatagcg taaagtatta tgggaggagt
      661 actaaagcgg acgccgtagt tgccgacctc agcgcacgca ataaactgtt caaacgtgaa
      721 cgagatgcta ttaaatctaa taatcatctc actgaaaaca atctatacat tagcgattat
      781 aagatgttaa ccttcgacgt gtttcgacca ttatttgatt ttgtaaacga aaaatattgt
      841 attattaaac ttccaacttt attcggtaga ggtgtaatcg atactatgag aatatattgt
      901 agtctcttta aaaatgttag actgctaaaa tgcgtaagcg atagctggtt gaaagatagc
      961 gccattatgg tggctagtga tgtttgtaaa aaaaatttgg atttatttat gtctcatgtt
     1021 aagtccgtca ctaagtcttc ttcttggaag gatgtgaaca gtgttcaatt tagtatttta
     1081 aacaatccag tggatacgga attcattaat aagttcttag agttttcgaa tagagtatac
     1141 gaagctctct attacgttca ctcgttgctt tattctagta tgacttctga ttcaaaaagt
     1201 atcgaaaaca aacatcagag aagactagtt aaactactgc tgtgaagatc ttcgaattca
     1261 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     1321 acaaatacca ctgagatcga tctttttccc tctgccaaaa attatgggga catcatgaag
     1381 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     1441 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     1501 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     1561 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     1621 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     1681 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     1741 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     1801 cccaagctct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     1861 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     1921 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     1981 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2041 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2101 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgcatgcc
     2161 tgcaggtcgg ccgccacgac cggtgccgcc accatcccct gacccacgcc cctgacccct
     2221 cacaaggaga cgaccttcca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     2281 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     2341 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     2401 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     2461 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     2521 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     2581 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     2641 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     2701 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     2761 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     2821 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     2881 acgaccccat ggctccgacc gaagccaccc ggggcggccc cgccgacccc gcacccgccc
     2941 ccgaggccca ccgactctag aggatcataa tcagccatac cacatttgta gaggttttac
     3001 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3061 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3181 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     3241 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3301 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3361 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3841 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4201 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4261 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4321 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4381 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4441 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4501 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4561 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4621 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4681 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4741 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4801 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4861 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4921 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4981 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5041 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5161 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt
     5221 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5281 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5341 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5401 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5461 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5521 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     5581 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     5641 tgtatttatt tattttttaa ttattt
//
